SU1406154A1 - Culture medium for cultivating balantidia - Google Patents

Abstract

The invention relates to biology, namely, nutrient media formulations, and can be used for the cultivation of parasitic ciliates. The aim of the invention is to improve the properties of the medium. To obtain the medium, 6.5-8.5 g of sodium chloride, 0.35-0.68 g of diammonium phosphate or 1.0-2.0 g of sodium phosphate disubstituted and 0.3-0.7 g are weighed potassium phosphate acid monosubstituted or potassium chloride. The weights are dissolved in 1000 ml of distilled water. The solution is sterilized. Rice starch is then added. Store medium in a domestic refrigerator for up to 50 days. Before sowing balantids, a biofunctional stimulator FIBS is added to the medium in the amount of 80-150 g / l. The reproduction of protozoa on the environment is already observed at the 12th hour after sowing. The maximum growth reaches 72-120 hours. The density of parasites is 7.8–9.0 thousand hours in 1 ml of medium. The survivability of balantidia in the proposed medium reaches 14 days. The medium is used for diagnostic purposes and for accumulation of pathogen biomass. 1 tab. with (L

Description

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ABOUT)

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The invention relates to biology, namely, nutrient media formulations, and can be used to cultivate infusoria.

The aim of the invention is to improve the properties of the medium.

The medium allows cultivating balantids of pigs without re-seeding and obtaining intensive accumulation of protozoa biomass.

The nutrient medium is prepared as follows.

An example. The optimal amounts of the components are weighed: 6.8 sodium chloride, 1.2 g phosphoric layer of disubstituted sodium, 0.5 g potassium chloride and dissolve them in 1000 MP of distilled water. The solution is sterilized by fusion for 5 minutes. After sterilization, the saline solution is filtered twice through filter paper and brought to volume with distilled water. The pH of the medium is measured and titrated to 7.2-8.0. The medium is poured into test tubes of 9 MP and sterilized in an autoclave. After cooling down, rice starch is added to the tip of the scalpel to each tube with the medium and left to complete bleaching. Store medium in a domestic refrigerator for up to 45 days.

Before sowing balantidia, FIBS 1 ml is added to each tube.

Into the obtained nutrient medium, the test material containing 300–900 copies of balantids is contained in 1 g of fecal masses. B quality control using a known medium. The results are presented in the table.

The number of trophozoites of balantidia in 1 ml of medium, th.

Offered

Known

1.8 2.2-3.1

1.5 2.0-2.5

Table continuation

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Reproduction of protozoa on the proposed medium is observed already by the 18th hour after sowing. The density of parasites reaches 1800 in 1 ml, and in a known medium - 1500.

Example 2. Weigh the optimal amounts of the components: 6.6 g of sodium chloride, 0.6 g of diammonium phosphate, 0.5 g of phosphoric acid, single-substituted potassium, and dissolve them in 1000 ml of distilled water. The solution is sterilized by boiling for 5 min. After sterilization, the salt solution is filtered twice through filter paper and the volume is brought to 1 l with distilled water. The pH of the medium is measured and titrated to 7.2-8.0. Wednesday poured into test tubes

9 ml and sterilized in an autoclave. After cooling down, rice starch is added to the tip of the scalpel to each tube with the medium and left to complete bleaching. Store medium in a domestic refrigerator for up to 50 days.

Before sowing balantidia, FIBS 1 ml is added to each tube.

The resulting nutrient medium is sown with material containing balantidium trophozoites in an amount of 300–900 copies per 1 g of fecal mass. As control use known medium.

Reproduction of protozoa on the proposed environment is noted by the 12th hour after sowing. The density of parasites reaches 2000 in 1 ml. The maximum growth is observed at 72-120 hours (up to 9000 trophozoites), while on the control medium there were a maximum of 5,000 ciliates per ml.

The first reseeding is carried out after 24 hours. Subsequent reseeding is carried out every 120 hours. At the same time, from the same amount of nutrient medium, the simplest biomass is obtained almost 2 times more than in the control. The survivability of balantidia in the resulting medium reaches 336 hours. The medium is suitable both for separating balantidia from feces, and for their further passage and accumulation.

Claims (1)

Invention Formula Nutrient medium for cultivation of balantids, containing mineral supplement, biogenic stimulator PhIBS, starch and distilled water, characterized in that, in order to improve the properties of the medium, as a mineral supplement contains sodium chloride, phosphoric acid, disubstituted sodium or hydrogen phosphate, disubstituted sodium, phosphoric acid, disubstituted sodium or a disubstituted disubstituted sodium, phosphoric acid, disubstituted sodium or a disubstituted disubstituted , potassium chloride or phosphoric acid monosubstituted potassium at the following quantitative ratio of components, g / l :. Sodium Chloride Phosphorus no acid 6.5-8.5 disubstituted ammonium or Phosphate disubstituted sodium Phosphoric acid monosodium potassium or Potassium chloride Biogenic stimulator Phibs Starch Distilled water